MOLECULAR BIOLOGY OF STEROID HORMONE ACTION

类固醇激素作用的分子生物学

• 批准号: 3273327
• 负责人: GORDON M RINGOLD
• 金额: $ 27.29万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-12-01 至 1987-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3273327
• 关键词: Escherichia coli; complementary DNA; cytochrome P450; endonuclease; eukaryote; flow cytometry; gene expression; genetic manipulation; genetic recombination; genetic transcription; glucocorticoids; hepatocellular carcinoma; hormone regulation /control mechanism; immunochemistry; liver; messenger RNA; microorganism genetics; molecular cloning; monoclonal antibody; mouse mammary tumor virus; nucleic acid sequence; radiotracer; spectrometry; steroid hormone; temperature sensitive mutant; vaccinia virus; viral carcinogenesis; virus RNA

Virtually all mammalian cells contain receptors for one or more classes of steroid hormones. These high-affinity receptor proteins most likely mediate the major effects of steroids on cell function by interacting with specific DNA sequences in (or near) target genes. Upon so doing, the steroid-receptor complex stimulates initiation of transcription thereby leading to increased production of specific RNAs. The study of complex regulatory mechanisms in higher eukaryotes has been hampered by the difficulty in manipulating such organisms genetically and by the enormity of mammalian genomes. We therefore have turned to simplified tissue culture systems and isolated genes to study the molecular mechanisms by which steroid hormones regulate gene expression. We will use genetic biochemical and molecular approaches to investigate the following issues: 1) What is the structure of the glucocorticoid receptor and which portions of the protein are required for its hormone binding, DNA binding, and transcription stimulating activities? 2) What are the biochemical components required for steroid-regulated transcription? 3) What factors determine whether a gene is or is not steroid responsive in a given cell (tissue) type? 4) How do steroid hormones alter the developmental pattern of gene expression in a well defined differentiating system? We will focus our attention on studies of the mouse glucocorticoid receptor and two specific genes that are regulated by glucocorticoids; one is the rat gene encoding the liver protein alpha-1 acid glycoprotein and the second is a mouse gene encoding a developmentally regulated cytochrome P450. It is our intent to eventually describe in detailed molecular terms the nature of the interactions between the glucocorticoid receptor and the transcriptional machinery of the cell. The approaches we are taking should help not only in elucidating the molecular mechanisms of steroid hormone action but may provide additional insights into the basis for tissue-specific gene expression and the process of cell differentiation.

几乎所有的哺乳动物细胞都含有一种或多种受体 类固醇激素的种类。这些高亲和力受体蛋白 很可能介导类固醇对细胞功能的主要影响 通过与靶标中(或附近)的特定DNA序列相互作用 基因。一旦这样做，类固醇-受体复合体就会刺激 启动转录，从而导致产量增加 特定的RNA。复杂调控机制的研究 在高等真核生物中的研究一直受到困难的阻碍 从基因上操纵这种有机体，并通过 哺乳动物的基因组。因此，我们转向了简化 组织培养系统和分离的基因来研究分子 类固醇激素调节基因表达的机制。 我们将使用遗传、生化和分子方法来 调查以下问题：1)是什么结构 糖皮质激素受体和蛋白的哪些部分是 它的激素结合、DNA结合和转录所需 刺激活动？2)生化成分是什么？ 类固醇调节转录所必需的？3)哪些因素 确定一个基因是否对给定的类固醇有反应 细胞(组织)类型？类固醇激素如何改变 基因表达的发育模式在一个明确的 差异化系统？ 我们将重点关注小鼠糖皮质激素的研究。 受体和两个受 糖皮质激素；一个是编码肝脏蛋白的大鼠基因 第一个是α-1酸性糖蛋白，第二个是小鼠编码基因 一个发育调节的细胞色素P450。我们的意图是 最终用详细的分子术语描述 糖皮质激素受体与血管内皮生长因子的相互作用 细胞的转录机制。我们正在采取的方法 服用应该不仅有助于阐明分子 类固醇激素的作用机制，但可能提供额外的 对组织特异性基因表达基础的洞察和 细胞分化的过程。